All issues
Volume 65 (1969)
- Issue 6 Pages 849-
- Issue 5 Pages 667-
- Issue 4 Pages 497-
- Issue 3 Pages 329-
- Issue 2 Pages 159-
- Issue 1 Pages 1-
Volume 65, Issue 2
Displaying 1-22 of 22 articles from this issue
- |<
- <
- 1
- >
- >|
-
SHOSUKE TAKEMURA, MASAZUMI MIYAZAKI1969 Volume 65 Issue 2 Pages 159-169
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSSeveral amino acid-specific transfer ribonucleic acid's (tRNA) were purified from Torulopsis utilis by column chromatography on DEAE-Sephadex. Digestion with pancreatic ribonuclease (RNase) followed by two-dimensional fractionation on chromatographic paper gave rise to a number of mono-and oligonucleotides. The distribution pattern of the digestion products was characteristic of each tRNA, indicating clear differences in the primary structure of the tRNA's. The 3'-terminal groups in all tRNA's were adenosine (1 mole per 75 to 90 moles of nucleotides). Many fragments from alanine-I and valine-I tRNA's were identified by the qualitative and quantitative analysis of their degradation products. Nucleotide sequences in these tRNA's were compared with those in the same specific tRNA's from baker's yeast. The two-dimensional procedure used here is very simple and suitable for rapid analysis of oligonucleotides, but is not so high in accuracy as to be used for determination of sequence of tRNA.View full abstractDownload PDF (2047K) -
FUMIO WADA, KAZUYA HIRATA, YUKIYA SAKAMOTO1969 Volume 65 Issue 2 Pages 171-175
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSParticipation of cytochrome P-450 in cholesterol synthesis by rat liver postmito-chondrial fractions was examined since cholesterol is synthesized by a series of reactions, including certain microsomal ones, which require both NADPH and molecular oxygen.
Cholesterol synthesis from various precursors, namely acetate, mevalonate or squalene, was inhibited by carbon monoxide and was accelerated with liver preparations from the phenobarbital-treated rats as compared with those from the untreated rats. This dif-ference in rate was much more significant with mevalonate as precursor than with acetate, and greater with squalene than with mevalonate. The addition to the reaction mixture of drugs, which were metabolized by enzyme systems involving cytochrome P-450, in-hibited the cholesterol synthesis. From these results, it is probable that cytochrome P-450 is functional in cholesterol synthesis.View full abstractDownload PDF (370K) -
SHOICHI NAKASHIMA, KIYOSHI TAKATSU, KINJI TSUKADA, HIKOKICHI OURA1969 Volume 65 Issue 2 Pages 177-183
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSAntibody formation was significantly enhanced by partial hepatectomy, performed 5 or 20 days after antigenic stimulation. On the other hand, little effect of partial hepatectomy on antibody formation was observed in rats operated 5 days before immunization or in sham-operated rats.
The incorporation of 3H-Ieucine into ribosomal fractions of splenic and lymph node cells was increased one day after partial hepatectomy. The incorporation of 3H-Ieucine into specific antibody molecule was elevated in two days after the operation.View full abstractDownload PDF (489K) -
V. Piece-Meal Hydrolysis of Native DNA Effected by DNase K2MIKIO KATO, SHUKUKO IKAWA, YONOSUKE IKEDA, MOTOHIRO FUKE1969 Volume 65 Issue 2 Pages 185-190
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSBacillus subtilis DNA was digested with DNase K2 [EC 3. 1. 4. 5] in the absence of MgSO4. Under the conditions used, the digestion proceeded slowly and the products consisted mainly of DNA segments with long chain lengths. After fractionation on a DEAE-cellulose column, the average chain length was estimated from the sizes of DNA appeared on electron-microscopic photographs. The DNA segments isolated from a fraction and purified by the gel filtration were found to be of comparatively homo-geneous size (500-1000 nucleotides in pairs).View full abstractDownload PDF (2481K) -
I. Purification, Properties and Application in Carbohydrate ResearchMINORU FUKUDA, TAKASHI MURAMATSU, FUJIO EGAMI1969 Volume 65 Issue 2 Pages 191-199
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSβ-Xylosidase [EC 3. 2. 1. 37] from the liver of Charonia lampas, a marine gastropod, was purified 89-fold using p-nitrophenyl β-D-xyloside as substrate.
Column chromatography with CM-Cellulose, Sephadex G-200 and hydroxylapatite and heat treatment were effective for its purification.
The purified β-xylosidase was practically free from proteinase activity and all other glycosidase activities tested except that of β-glucosidase [EC 3. 2. 1. 21].
The enzyme released xylose from xylosyl protein, which is considered to be a pre-cursor to chondromucoprotein. It also hydrolyzed O-β-xylosyl-L-serine, xylopentaose and xylan.View full abstractDownload PDF (639K) -
ASAKO YUTANI, KATSUHIDE YUTANI, TOSHIZO ISEMURA1969 Volume 65 Issue 2 Pages 201-208
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe molecular weight of saccharifying bacterial a-amylase [EC 3. 2. 1. 1] was deter-mined to be 4.1×104 by Archibald method. The sedimentation coefficient was 4.02S and the value of intrinsic viscosity was 0.034 dl per gram. The contents of tyrosine and tryptophan residues were determined spectrophotometrically, and found to be 13 and 11 moles per mole of protein, respectively. About 80 per cent of the tyrosine residues were abnormal in ionization, and about 80 per cent of the tryptophan residues seemed to be on the surface of the molecule. The measurements of optical rotatory dispersion and circular dichroism indicated that the helical content of this protein was about 20 percent. The enzyme was inactivated with 8M urea containing EDTA or HQ at 37°C. In these state, the secondary and tertiary structure of native enzyme was completely destroyed. The denatured enzyme was reactivated when it was diluted with the buffer containing calcium. The calcium ion was necessary for the stability of the conformation and the refolding of the unfolded polypeptide to the correct confor-mation.View full abstractDownload PDF (520K) -
XX. Biochemical Studies. (ll). Enzymatic Conversion of 2'-Ketopantothenate to D-Pantothenate in RatYASUSHI ABIKO, MUNEHIRO TOMIKAWA, MASAO SHIMIZU1969 Volume 65 Issue 2 Pages 209-214
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThe in vivo and in vitro conversion of 2'-ketopantothenate to a-pantothenate was demonstrated in rat. The enzyme which catalyzed the reductive conversion was localized in the microsomal and the soluble fractions of rat liver, and both enzymes required NADH or NADPH as a cofactor. Two distinct enzyme systems were found in the microsomal fraction. One was NADH-dependent and the other NADPH-depen-dent. The NADPH-dependent activity was stimulated in an anaerobic condition and increased by phenobarbital treatment of rat. The soluble enzyme was different from the microsomal enzymes in pH optimum, the effect of the substrate concentration and thermostability.View full abstractDownload PDF (435K) -
NAGASUMI YAGO, SHOGO ICHII1969 Volume 65 Issue 2 Pages 215-224
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Osmotic shock followed by discontinuous Ficoll gradient centrifugation was applied to hog adrenocortical mitochondria to localize components of the steroid 11β-hydroxylase [EC 1. 14. 1.6] system and of the cholesterol sidechain-cleaving enzyme system in submitochondrial fractions.
2. The supernatant obtained by centrifugation at 17, 000×g after the osmotic shock at 0°C in hypotonic phosphate buffer, contained high malate dehydrogenase [EC 1. 1. 1. 37] activity. Neither the steroid 11β-hydroxylase system nor the cholesterol sidechain-cleaving enzyme system was found in this fraction.
3. Of the four fractions obtained by discontinuous Ficoll Gradient centrifugation after osmotic shock, the uppermost fraction was judged to contain the outer membranes of mitochondria from its contents of monoamine oxidase [EC 1. 4. 3. 4], Amytal-insensi-tive NADH cytochrome c reductase and a cytochrome tentatively identified as cyto-chrome b5. A significant amount of antimycin A-insensitive NADPH cytochrome c reductase was also found in this fraction.
4. The pellet and two other membranous fractions obtained by discontinuous Ficoll gradient centrifugation seemed to contain materials derived from the inner membranes of mitochondria. NADPH diaphorase [EC 1. 6. 99. 1], cytochrome P-450, steroid 11β-hydroxylase and enzymes which participate in the reaction cleaving the cholesterol sidechain, i.e., 20α-hydroxylase, 22-hydroxylase and C20, 22-lyase, could be localized in the inner membrane but not in the outer membrane. Adrenodoxin seemed to leak out of these enzyme complexes of steroid hydroxylation during preparation of submitochondrial fractions.View full abstractDownload PDF (766K) -
YOSHIO IMAI, KAZUKO IMAI, RYO SATO, TAKEKAZU HORIO1969 Volume 65 Issue 2 Pages 225-237
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. Oxidized cytochrome cc' from Rhodospirillum rubrum exhibits three different types of absorption spectrum depending on pH; i.e., the neutral type (Type I), intermediate type (Type II), and alkaline type (Type III). The pH-dependent spectral conversion is reversible. In the reduced form, it shows an abnormal spectrum (Type α) in the pH region where the spectrum of the oxidized form is either Type I or II, but is converted to a state having a normal spectrum (Type n) at more alkaline pH's.
2. Alcohols, phenols and ketones induce the spectral conversion from Types I and II to Type III at fixed pH values, and the efficiency of these solvents in causing the conversion parallels their hydrophobicity. Urea and guanidine hydrochloride also effect the conversion at slow rates. On the other hand, high ionic strength not only prevents the conversion of Type II to Type III, but also causes almost complete reversion of Type III to Type II at alkaline pH's. Glycerol shows the same effect to a lesser extent. Deoxycholate shows no effects.
3. The helical content determined from circular dichroism measurements is about 63% in both the states showing Type I and II spectra. The spectral conversion by 2-propanol at pH 7 results in no appreciable change in the helical content, whereas that induced by alkali or urea is accompanied by a considerable increase in the random-coli structure.
4. Cytochrome c' from Rhodopseudomonas palustris responds to pH and the various reagents similarly, but is more resistant to treatments causing the conversion to Type III.
5. It is concluded that the anomalous Type I and II spectra of these cytochromes arise from a unique hydrophobic structure in the vicinity of the heme. Direct and indirect destruction of this structure leads to the conversion to Type III spectrum, which is normal for c-type cytochromes. This conversion does not necessarily involve the alteration in the gross helical conformation of the proteins.
6. These properties of cytochromes cc' and c' are discussed in relation to the atypical spectral properties of P-450 and the ethyl isocyanide compound of protoheme.View full abstractDownload PDF (989K) -
III. Precipitation Reaction of the Low Density Lipoprotein Fraction of Sera of Various Animals with Synthetic Glycosphingosyl-Protein ConjugatesTAMOTSU TAKETOMI1969 Volume 65 Issue 2 Pages 239-246
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIt was found that glycosphingosyl-protein conjugates precipitate with whole sera of various animals, particularly with their low density lipoprotein fractions in agar gel double diffusion. Among the sera tested, human serum showed the strongest band with glycosphingosyl-protein conjugates. It was noted that the quantitative precipitation reactions between the low density lipoprotein fractions of various sera and the galactosyl-sphingosyl-protein conjugates were just like the quantitative precipitation reaction.View full abstractDownload PDF (2827K) -
KOITI TITANI, KOZO NARITA1969 Volume 65 Issue 2 Pages 247-257
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSIn order to elucidate the complete amino acid sequence of baker's yeast (Saccharomyces oviformis) cytochrome c, isolation and sequence determination of peptic peptides were performed. Only one (P-XXVIII-2) out of thirty-one peptides isolated provided direct evidence for the linkage between two of the four peptide fragments, whose arrangement had remained undetermined in the study of tryptic and chymotryptic peptides. Other peptides provided confirmatory data for the proposed amino acid sequence within each fragment.View full abstractDownload PDF (663K) -
KOZO NARITA, KOITI TITANI1969 Volume 65 Issue 2 Pages 259-267
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSThrough the studies of the chymotryptic and peptic peptides derived from cytochrome c of baker's yeast (Saccharomyces oviformis), twenty-two tryptic peptides could be combined into five large peptide fragments, but the mutual arrangement of the above five fragments remained to be determined. In the present paper, two approaches were attempted in order to know the order of the five fragments. The one was the cleavage of the two methionyl residues in the protein to isolate the expected three fragments. These fragments were analyzed and the complete amino acid sequence of the protein containing 108 amino acid residues were established. Another approach was to cleave three arginyl bonds with trypsin of the succinylated cytochrome. By analyzing the amino acid compositions of the four tryptic peptide fragments, the arrangement of the above five fragments could be determined and the same conclusion as that deduced from the cyanogen bromide experiment has been drawn.View full abstractDownload PDF (1171K) -
V. On the Substrate-Induced Spectral Change of P-450 Solubilized from Bovine Adrenocortical MitochondriaFUMIKO MITANI, SHIGEO HORIE1969 Volume 65 Issue 2 Pages 269-280
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSProcedures for solubilization and purification of P-450 from bovine adrenocortical mitochondria were examined and an improved method for the preparation of partially purified P-450 was described.
By measuring the “substrate-induced difference spectrum”, the substrate-binding specificity of the solubilized preparation was compared with those of P-450 in bovine adrenocortical mitochondria, bovine adrenocortical microsomes, and rat liver microsomes. The specificity of the solubilized preparation was mostly the same as that of P-450 in adrenocortical mitochondria. The specificity of mitochondrial P-450 and that of P-450 in adrenocortical microsomes were similar in qualitative aspects, but the relative intensity of spectral change and the affinity for substrate were frequently different. On the other hand, the binding specificity of hepatic microsomal P-450 was rather different qualitatively from those of adrenal P-450's.
The effect of successive addition of two different substrates to the solubilized prepa-ration was also examined. The intensity of spectral change induced by a type I sub-strate was inhibited non-competitively by the addition of another type I substrate, but was not affected by the addition of a type II substrate. The significance of these results was discussed.
In addition to these observations, spectral evidence was presented indicating the change in spin state of P-450 by binding with a substrate.View full abstractDownload PDF (1619K) -
KAZUHIKO KONISHI, TAKASHI TAKAGI1969 Volume 65 Issue 2 Pages 281-284
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. The fibrin stabilizing factor (FSF) was purified by stepwise elution from a DEAE-cellulose column with Tris and was activated by trypsin [EC 3. 4. 4. 4].
2. Activated FSF showed an autocatalytic activation and activated FSF increased the turbidity of fibrinogen.
3. These results indicate that activated FSF has a proteolytic action and the activa-tion of FSF involves limited proteolysis.View full abstractDownload PDF (237K) -
VIII. Intermediate Formation and Activation of Myosin by ATPNAOKAZU KINOSHITA, SHUICHIRO KUBO, HIROFUMI ONISHI, YUJI TONOMURA1969 Volume 65 Issue 2 Pages 285-301
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESS1. A stable NTP-myosin compound, which was produced by incubating myosin ATPase [ATP: phosphohydrolase, EC 3. 6. 1. 3] with NTP in the presence of ATP and Mg++, was isolated by gel-filtration through a Sephadex column. The amount of initial burst of Pi-liberation from the myosin-ATP system decreased almost linearly with increase of bound NTP, and was reduced to zero on binding about I mole of NTP per 4×105g of myosin.
2. The maximum velocity, Vmax, and the pH-activity curve of myosin ATPase in the steady state were scarcely changed on the stable binding of l mole of NTP to 4×105g of myosin. However, the value of the Michaelis constant, Km, decreased on binding of NTP to myosin.
3. Analysis of the ultraviolet absorption spectrum of NTP bound to myosin indicated that NTP combines with a residue in the myosin molecule through its sulfhydryl group. To determine the amino acid residue with which NTP combines, NTP-myosin was digested by Nagarse [Subtilopeptidase A; EC 3. 4. 4. 16] and Pronase [EC class 3. 4. 4], and the NTP-peptide mixture was isolated from the digest by chromatography on Amberlite IRC-50 resin. The bond between NTP and the peptide of the NTP-peptide mixture thus obtained was found to be stable at acidic and neutral pH values, but unstable at alkaline pH's. NTP-glutamic acid was isolated almost quantitatively by column chromatography from the complete digest of the NTP-peptide mixture by Ieucine aminopeptidase [EC 3. 4. 1. 1] and prolidase [EC 3. 4. 3. 7].
4. The velocity of liberation of free ADP after addition of 1 mole of ATP to 3.4×105g of H-meromyosin was determined by measuring the velocity of pyruvate-liberation on addition of pyruvate kinase [ATP: pyruvate phosphotransferase, EC 2. 7. 1. 40] and PEP at appropriate times after the start of the reaction. The rate of ADP-liberation P from the myosin-phosphate-ADP complex, _??_, was estimated by analysis of the ADP results. It was much lower than the rate of formation of this complex and far higher than the rate of decomposition of this complex to E+ADP+Pi+H+.
5. The report by Kanazawa and Tonomura on the existence of an extra-burst of Pi liberation in the initial phase of the reaction of myosin and ATP at a low concen-tration of Mg++ and high concentration of KCl at room temperature was confirmed. The amount of the extra-burst of Pi-liberation increased with increase in Mg++ concen-tration from 0 to 10μM, and reached a maximum level of 10-20 moles Pi per 4×105g of myosin at 10μM Mg++. It decreased gradually with further increase in Mg++ concen-tration. In the presence of more than 1mM Mg++, the initial burst remained at a stoichiometric amount of 1 mole Pi per 4×105g of myosin. The effects of temperature, KCl concentration and treatment with PCMB-β-mercaptoethanol on the amount of the extra-burst of P liberation were also examined.
6. The time-courses of TCA labile Pi and hydrogen ion-liberation from the myosin-ATP system were measured in the presence of 10μM Mg++. The extra-burst of TCA-labile Pi-liberation amounted to 17 moles per 4×105g of myosin and that of hydrogen ion-liberation to 10.5 moles per 4×105g of myosin. The initial rapid liberation of ADP from the myosin-ATP system during the extra-burst of Pi-liberation was followed by measuring pyruvate-liberation in a reaction medium coupled with the pyruvate kinase system. The initial rapid ADP-liberation was about I mole less than the amount of the initial burst of P liberation per 4 ×105g of myosin. These results indicate that during the extra-burstView full abstractDownload PDF (1763K) -
ATSUSHI KIMURA, KOICHI TSURUMI1969 Volume 65 Issue 2 Pages 303-304
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (1252K) -
TATSUYA SAMEJIMA, MASAKO KITA, MASACHIKA IRIE1969 Volume 65 Issue 2 Pages 305-307
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (196K) -
AKIRA IMANISHI, TOSHIZO ISEMURA1969 Volume 65 Issue 2 Pages 309-312
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (288K) -
YUKIHIKO SAKURAI, TARO HAYAKAWA, YUKIKO FUKUYOSHI, MASAHIKO KOIKE1969 Volume 65 Issue 2 Pages 313-315
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (822K) -
TAKAYUKI OZAWA1969 Volume 65 Issue 2 Pages 317-319
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (205K) -
KENSAKU MORIHARA, KENZO NAGAMI1969 Volume 65 Issue 2 Pages 321-323
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (215K) -
ATSUSHI TAKAHASHI, RYUICHI KATO1969 Volume 65 Issue 2 Pages 325-327
Published: February 25, 1969
Released on J-STAGE: November 18, 2008
JOURNAL FREE ACCESSDownload PDF (194K)
- |<
- <
- 1
- >
- >|